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Objective:To explore the clinical outcomes of top-quality blastocysts transfer developed from cleavage embryos with different grading and determine whether the cleavage stage embryo morphology grading should be taken into consideration when transferring the embryo at the blastocyst stage.Methods:A number of 3 059 cycles were included with single top-quality blastocyst transfer dating from January 2017 to May 2021 in Henan Provincial People′s Hospital. According to the number of cleavage sphere and degree of fragmentation, all cleavage stage embryos were divided into three groups: top D3 embryo (8 cells, ≤5% fragments)-TB group, suboptimal D3 embryo (8 cells, 5%<fragments≤10%; 7 cells or 9 cells, ≤10%)-TB group, and normal D3 embryo-TB group. Univariate analysis, multivariate logistic regression analysis and threshold effect analysis were performed on the data.Results:The clinical pregnancy rates of top D3 embryo-TB group(1 326 cycles), suboptimal D3 embryo-TB group (830 cycles) and normal D3 embryo-TB group (903 cycles) were 69.53%, 70.12% and 66.67%, respectively ( P>0.05); and the early abortion rate were 10.74%, 12.54% and 12.62%, respectively ( P>0.05). After adjusting for confounders, logistic regression showed that no significant associations were found between cleavage stage embryo morphology grading and clinical pregnancy rate (suboptimal D3 embryo-TB group: OR=1.02, 95% CI: 0.76-1.38, P=0.879; normal D3 embryo-TB group: OR=0.84, 95% CI: 0.61-1.14, P=0.262) and early abortion rate (suboptimal D3 embryo-TB group: OR=1.18, 95% CI: 0.77-1.82, P=0.445; normal D3 embryo-TB group: OR=1.26, 95% CI: 0.81-1.98, P=0.309). The results of threshold effect analysis showed that when a single top-quality blastocysts was transferred, the effect of age on the clinical pregnancy rate showed a curve relationship, when the age was≥33 years old, the clinical pregnancy rate decreased significantly with age increased ( OR=0.89, 95% CI: 0.83-0.95, P=0.007); and there was no significant change in early abortion rate ( OR=1.01, 95% CI: 0.97-1.06, P=0.628). Conclusions:Cleavage stage embryo grading is not found to correlate with clinical outcomes in single top-quality blastcyst tranfer. Therefore, when considering blastocyst transfer, its morphology at blastocyst stage is more relevant. The effect of age on pregnancy outcomes of single blastocyst transfer should be considered.
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Objective To explore the relationship between clinical outcomes and embryo transfer number, and to provide some proposals for transfer selection of elderly patients. Methods Data from 80 fresh transfer cycles with cleavage-stage embryos were analyzed. Cycles were divided into several groups according to transfer number. Clinical pregnancy rate, implantation rate, multiple pregnancy rate and live birth rate were compared. Results To women no younger than 38 years old, when available embryo number was larger than two, similar clinical outcomes could be achieved by transferring two and three embryos. This trend was independent of the number of transferred 8-cell embryos. Conclusion The number of fresh cleavage-stage embryos transfer should not exceed two in elderly women.
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@#【Objective】To investigate the impact of long- term storage time on epigenetic modification of histone in human cleavage stage embryos.【Methods】According to the length of storage time，donated embryos after slow-freezing were divided into 3 groups ：6-year group ，9-year group ，and 12-year group ，while the control group consisted of donated fresh embryos. Immunocytochemistry was performed to compare the expression levels of HDAC1， H3K9ac， H3K4me3 ，and H3K9me3 among 4 groups. Transcription levels of HDAC1 ，SUV39H1 ，SETDB1 ，and KDM5A were analyzed through Single-Cell qRT-PCR.【Results】The relative abundances of HDAC1 and SUV39H1 mRNA showed no significant differences among 4 groups（P > 0.05）. SETDB1 exhibited a climbing pattern as storage time increased，but no significant difference was observed（P > 0.05）. There were no differences in H3K9 trimethylation and H3K9 methylation among 4 groups. However ，the expression level of KDM5A increased with the increasing storage time（P < 0.05）.【Conclusions】 Storage time did not affect the expression of deacetylase HDAC1，methylase SUV39H1 and SETDB1. H3K9ac/me3 and H3K4me3 also exhibited no significant difference as the storage time increases. However ，the increasing storage length might induce the elevating expression of demethylase KDM5A，which may be associated with inhibition of embryonic transcription.
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[Objective]To compare the single live birth outcomes of blastocyst transfer between vitrified blastocyst and blastocyst cultured from thawing cleavage embryo,so as to choose the best scheme of blastocyst transfer.[Methods]Retrospective analysis of the single live birth clinical data of 1 037 vitrified blastocyst compared with 690 blastocyst cul-tured from thawing cleavage embryo undergoing frozen embryo transplantation(FET)from January 2014 to October 2016 was performed.Mail outcome were including gestational age,neonatal weight,proportion of male neonate,preterm birth rate,very preterm birth rate,low birthweight rate,very low birthweight rate,congenital anomalies rate.[Results]There were no differences between the two groups for gestational age,neonatal weight,proportion of Live birth,health baby and stillbirth(P>0.05). There were no differences in proportion of male neonate(AOR 1.07,95% CI 0.86~1.34),preterm birth rate(AOR 0.7,95% CI 0.49~1.01),very preterm birth rate(AOR 1.47,95% CI 0.55~3.96),low birthweight rate (AOR 1.38,95% CI 0.86~2.22),very low birthweight rate(AOR 0.76,95% CI 0.20~2.83),congenital anomalies rate (AOR 1.58,95% CI 0.66~3.76,P>0.05).[Conclusion]The blastocyst may be the preferable stage for vitrifying and transfer currently which can obtain good neonatal outcomes.
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Objective To evaluate the efficiency of the application of array comparative genomic hybridization (array-CGH) in preimplantation genetic diagnosis or screening (PGD/PGS), and compare the clinical outcomes of different stage embryo biopsy. Methods The outcomes of 381 PGD/PGS cycles referred in the First Affiliated Hospital of Nanjing Medical University from July 2011 to August 2015 were retrospectively analyzed. There were 320 PGD cycles with 156 cleavage-stage-biopsy cycles and 164 trophectoderm-biopsy cycles, 61 PGS cycles with 23 cleavage-stage-biopsy cycles and 38 trophectoderm-biopsy cycles.Chromosomal analysis was performed by array-CGH technology combined with whole genome amplification.Single embryo transfer was performed in all transfer cycles.Live birth rate was calculated as the main clinical outcomes. Results The embryo diagnosis rate of PGD/PGS by array-CGH were 96.9%-99.1%. In PGD biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were 50.0%(58/116) and 37.2%(58/156) in cleavage-stage-biopsy group, 67.5%(85/126) and 51.8%(85/164) in trophectoderm-biopsy group (both P<0.01). In PGS biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were the same as 34.8%(8/23) in cleavage-stage-biopsy group, the same as 42.1%(16/38) in trophectoderm-biopsy group (both P>0.05). Conclusions High diagnosis rate and idea live birth rate are achieved in PGD/PGS cycles based on array-CGH technology.The live birth rate of trophectoderm-biopsy group is significantly higher than that of cleavage-stage-biopsy group in PGD cycles;the efficiency of trophectoderm-biopsy is better.
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OBJECTIVE: To determine the incidence of embryo retention (ER) in the transfer catheter following embryo transfer (ET) in blastocyst transfer and investigate whether retransferring retained embryos has an impact on reproductive outcomes in patients undergoing in vitro fertilization-ET. METHODS: We retrospectively analyzed the records of 1,131 blastocyst transfers, which comprised 223 single blastocyst transfer (SBT) and 908 double blastocyst transfer (DBT) cycles. Each SBT and DBT group was classified depending on whether ET was performed without retained embryos in the catheter during the first attempt (without-ER group) or whether any retained embryos were found following ET (ER group) for the purpose of comparing reproductive outcomes in a homogenous population. RESULTS: The overall incidence of finding retained embryos was 2.8% (32/1,131). There were no retained embryos in SBT cycles. In DBT cycles, implantation rates (30.0% vs. 26.6%), positive β-hCG rates (57.2% vs. 56.2%), clinical pregnancy rates (45.3% vs. 46.9%), and live birth rates (38.9% vs. 43.8%) were not significantly different between the without-ER and ER groups. There were no significant differences in the mean birth weight (g) 2,928.4±631.8 vs. 2,948.7±497.8 and the mean gestational age at birth (269.3±17.2 days vs. 264.2±25.7 days). A total of nine cases of congenital birth defects were found in this study population. Eight were observed in the without-ER group and one in the ER group. CONCLUSION: Our results suggest that retransfer of retained embryos does not have any adverse impact on reproductive outcomes in blastocyst transfer cycles. Furthermore, our results support finding that SBT might be advantageous for decreasing the incidence of retained embryos in catheters.
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Humans , Birth Weight , Blastocyst , Catheters , Congenital Abnormalities , Embryo Transfer , Embryonic Structures , Fertilization in Vitro , Gestational Age , In Vitro Techniques , Incidence , Live Birth , Parturition , Pregnancy Rate , Retrospective StudiesABSTRACT
Background: The objective of this retrospective study was to compare the efficacy of slow freezing and Vitrification for the cryopreservation of supernumerary cleavage stage embryos on day 3 after IVF and its impact on clinical outcome. Methods: 485 supernumerary embryos of IVF cycles (from Oct 2011 to Dec 2012) were cryopreserved by slow freezing method while 502 embryos (from Jan 2013 to April 2014) by Vitrification method. 362/485 and 230/502 embryos were thawed for FET cycles (65 patients in each group).After warming the survival rate, post warmed embryo morphology, clinical pregnancy and implantation rates were evaluated and compared between the two groups. Results: There were 65 frozen thawed cycles in each group. The percentage of excellent and good morphology embryos before cryopreservation were same in both the groups, but after thawing the results were significantly in favour of Vitrification as compared to Slow freezing. In Vitrification group versus Slow freezing group, the different outcomes were survival rate (96.95% vs. 69.06%, p-0.000), post warmed excellent morphology embryos (94.17% vs. 60.8%, p-0.000) clinical pregnancy rate (41.53% vs. 21.53%, p-0.043) and the implantation rate (14.41% vs. 7.01%, p-0.024). Conclusions: Vitrification is a promising alternate to the conventional slow freezing method in terms of not only excellent survival and post warmed excellent morphology embryo rate but also higher clinical pregnancy and implantation rate.
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Objective To analyze the outcomes of frozen-thawed blastocyst and cleavage embryo transfer after whole embryos cryopreservation. Methods The data of 489 IVF-ET cycles in reproductive medicine center of our hospital from September 2012 to August 2013 were analyzed retrospectively. Whole embryos cryopreservation in 214 patients were carried out with vitrification method and served as group A , 275 cycles performed fresh embryo transfer were served as group B. Then group A and group B were subdivided into group A1 (83 cycles),group A2 (131 cycles), group B1 (120 cycles)and group B2 (155 cycles)according to blastocyst transfer or cleavage-stage embryo transfer. The clinical outcomes of all groups were compared each other. Results The pregnancy rate and embryo implantation rate in group A1 were significantly higher(71.1% ,53.0%)than those in group A2, B1 and B2 (A2 group: 57.3%, 34.0%, B1group: 55.0%,42.1%, B2 group: 52.9%, 32.7%,respectively)(P<0.05). The embryo implantation rate in group B1 were higher than those in group B2 (P < 0.05). Conclusion F-ET after whole embryos freezing could significantly improve the embryo utilization rate and clinical pregnancy rate. Frozen-thawed blastocyst transfer could get better clinical outcomes than frozen-thawed cleavage-stage embryo transfer.
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PURPOSE: Blastocyst transfer has been recommended to raise the implantation rate without affecting the pregnancy rate. The objective of this meta-analysis is to systematically evaluate whether the live birth rate and other pregnancy outcomes can be improved by blastocyst transfer compared with cleavage-stage embryos transfer. MATERIALS AND METHODS: EMBASE and MEDLINE databases were searched for papers published between March 2004 and March 2013. An extensive range of the electronic databases yielded initially 317 studies from which seven trials met the inclusion criteria for further analysis. Our outcome measures were the live birth rate, clinical pregnancy rate, implantation rate, ongoing pregnancy rate, multiple pregnancy rate, first trimester miscarriage rate and ectopic pregnancy rate. Fixed effects models were chosen to calculate the odds ratio (OR). RESULTS: Seven trials (n=1446 cases) were finally analyzed. Compared with cleavage-stage embryos transfer, the blastocyst transfer was statistically significantly associated with an increase in clinical pregnancy rate [OR 1.43; 95% confidence interval (CI), 1.15-1.78], implantation rate (OR 1.38; 95% CI, 1.09-1.74) and ongoing pregnancy rate (OR 2.15; 95% CI, 1.57-2.94), and also a reduction in the probability of first trimester miscarriage rate (OR 0.51; 95% CI, 0.30-0.87). The improvement in the live birth rate was also observed (OR 1.77; 95% CI, 1.32-2.37). Moreover, there was no evidence of difference in multiple pregnancy and ectopic pregnancy rates. CONCLUSION: The available evidences suggest that live birth and other pregnancy outcomes after fresh in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) are significantly improved following blastocyst transfer as compared to cleavage-stage embryo transfer.
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Female , Humans , Pregnancy , Birth Rate , Embryo Transfer , Fertilization in Vitro/methods , Sperm Injections, IntracytoplasmicABSTRACT
Introdução: a biópsia embrionária tem como objetivo selecionar embriões geneticamente normais. Essa seleção ocorre por meio de testes genéticos pré-implantacionais. Espera-se, com isso, uma diminuição dos riscos de doenças genéticas e um aumento das taxas de implantação em fertilização in vitro. Objetivo: verificar, por meio de revisão bibliográfica, qual técnica de biópsia embrionária é considerada mais apropriada para feitura de testes genéticos pré-implantacionais. Método: pesquisa bibliográfica, na forma de revisão de publicações científicas, por meio das redes US National Library of Medicine (Pubmed), Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs), Google Acadêmico e Biblioteca Virtual em Saúde (BVS). Resultados e conclusão: existem três maneiras de efetuar a biópsia para reprodução humana assistida. A primeira consiste em retirar o primeiro e/ou o segundo corpúsculo polar estruído pelo oócito. Também se pode fazer a biópsia a partir de um blastômero do embrião em estágio de clivagem ou usar cinco a dez células do trofoectoderma de blastocisto. Normalmente as técnicas usadas para o diagnóstico são PCR, Fish, CGH array e SNP array, entre outras. Acredita-se que a biópsia de blastocistos é a melhor técnica para manter o potencial de implantação embrionária. Essa tendência se justifica por causa da maior quantidade de material genético disponível em fase avançada de desenvolvimento embrionário. Admite-se que nessa fase a incidência de mosaicismo seja menor em relação à biópsia de blastômeros, com consequente aumento na eficácia dos testes genéticos. Outra questão importante é que na biópsia de blastocistos as células são retiradas do trofoectoderma, enquanto que na biópsia em estágio de clivagem a remoção de um blastômero pode prejudicar o desenvolvimento embrionário.
Introduction: the embryo biopsy aims to select genetically normal embryos. This selection occurs through pre- implantation genetic testing. It is expected the reduction of risk ofgenetic disorders and increase implantation rates in IVF.Objective: to verify, through bibliographical revision, which embryo biopsy technique is considered more suitable for pre-implantation genetic diagnosis. Method: bibliographical research, in the form of literary review of scientific publications via networks, US National Library of Medicine (Pubmed), Latin-American Literature and Caribbean Health Sciences (Lilacs), Google Scholar and Virtual Health Library. Results and conclusion: there are three ways to perform the biopsy on assisted human reproduction.The first one consists in removing the 1st and/or 2nd polar body (if there wasfertilization). You can also perform the biopsy from the one blastomere of embryo cleavage stage or use 5-10 trophoectoderm cells blastocyst. Usually the techniques used for diagnosticpurpose are PCR, Fish, CGH array, SNP array and others. Nowadays it is believed that blastocyst biopsy is the best technique in order to maintain the embryonic implantation. This tendency is justified by the larger amount of genetic material available in an advancedstage of embryonic development. It is assumed that in this stage the incidence of mosaicism is reduced with the consequent increase in the effectiveness of genetic testing. Another important question is that the blastocyst biopsy cells are removed from the trophoectoderm while inbiopsy incleavage stage, the removal of one blastomere can impair embryonicdevelopment.
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Humans , Biopsy/methods , Choice Behavior , Embryo, Mammalian/cytology , Genetic Testing/methods , Blastocyst/cytology , Blastomeres/cytology , Cleavage Stage, Ovum , Embryo, Mammalian/pathology , Embryo Implantation/physiologyABSTRACT
This study was carried out to determine the effect of the duration of exposure to the infrared 1.48 mm diode laser, on the developmental potential of cleavage stage embryos. A total of 69 mouse embryos were included in the study. Twenty-two of which (group A) were biopsied using the laser with a longer duration of exposure (600 ms), meanwhile 47 (group B) were biopsied using the same laser with a shorter period (5 ms). The blastocyst formation rate of group B (46/47, 97.8%) was significantly higher than that of group A (12/22, 54.4%). There were no grade 1 blastocysts or hatching in group A. In contrast, 35 of 46 (76.0%) blastocysts in group B were grade 1 and the hatching rate of group B was 84.7% (39/46). In conclusion, the infrared 1.48 mm diode laser may be effective and safe with cautious application. A long duration of exposure to the laser can adversely affect the developmental potential of the biopsied embryos. The laser system with a shorter duration of exposure, therefore, is recommended for laser assisted embryo biopsy.
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Objective To evaluate the effects of biopsy methods, biopsy timing and the number of cell removed on in vitro development of embryos Methods One hundred and fifty four embryos of good morphology from in vitro fertilization patients were studied Sixty six embryos were allocated to the following three groups: chemical drilling biopsy group (26), mechanical drilling biopsy group (26) and control group (20) One cell was removed from the embryos of the two biopsy groups The remaining 88 embryos were allocated to two groups: biopsy group (44) and control group (44) Two cells were removed from biopsy group by chemical drilling method The stage of the embryo before biopsy, biopsy time, lysed blastomere, growth potential and hatching capacity of the biopsied embryos, total cell number at the blastocyst stage were recorded and evaluated Results The mean time of biopsy in the chemical drilling group (231?20) seconds was significantly shorter than that in the mechanical drilling group (262?23) seconds ( P 0 05) However, the proportion developing to the blastocyst stage was reduced after the removal of two cells from the 6 cell stage in comparison to the control (1/8 versus 5/8, P
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Objective To compare the effects of vitrification with slow-freezing on the developmental ability of day 3 cleavage stage embryos.Methods Patients who had no less than 4 high quality embryos were included in this study.These embryos were cryopreserved using the methods of vitrification or slow-freezing.In the eryopreserved embryo transfer cycles,the embryos which were cryopreserved using one of the methods were chosen randomly.The developmental ability of embryos was compared between these two groups.Results A total of 80 patients were included in this study with 160 embryos.In the group of slow-freezing,73(91%)embryos were survived and achieved 15(38%)clinical pregnancies.Among these,3 were twins and the implantation rate was 25%(18/73).In the group of vitrification,71(89%)embryos were survived and achieved 19(48%)clinical pregnancies.Among these, 9 were twins and the implantation rate was 39%(28/71),which was significantly higher than the slow- freezing group(P
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Objective To determine whether cleavage developmentally retarded embryos have not cleaved during a 24 hour period could develop into blastocysts and produce hESC cell lines.Methods A total of 120 such embryos were cultured to blastocyst stage by sequential culture.Blastocysts formation rate and quality of blastocyst were detected under microscope.The relation between blastocyst formation rate and blastomere number,the fragment of blastomere and blastomere symmetry were analyzed by stepwise Logistical regression analysis.Inner cell masses(ICMs)were isolated by immunosurgery.Colonies derived from the ICMs were passed every 4-7 days and the derivatives were passaged and identified.Results A total of 22 blastocysts were obtained from 120 embryos.The blastulation rate was 18.7%.Early blatocyst, blastocyst,full blastocyst,expanded blastocyst,hatching blastoeyst and hatched blastocyst accounted for 5.9%,23.5%,35.3%,23.5%,5.9%,and 5.9% respectively.The grade of ICM and trophoblast was mostly scored C or B.Blastocyst formation rate was related to cell number and blastomere symmetry but not fragment.Immunosurgery resulted in the formation of 7 ICMs and 3 primary colonies,which produced 2 cell lines.The cell lines satisfied the criteria that characterize pluripotent hESC cells.Undifferentiated cells were positive for AKP,SSEA-4,TRA-1-60,and TRA-1-81.It could continue to proliferate in vitro and form embryoid bodies when cultured in suspension.It had capability to form teratoma in SCID mice.Both cell lines had normal karyotypes after 45 and 34 passages respectively.Conclusions Our results suggest that a subset of developmentally retarded embryos can form blastocysts and give rise to hESC cell lines.